title: Supervillin binding to myosin II and synergism with anillin are required for cytokinesis.
abstract: Cytokinesis, the process by which cytoplasm is apportioned between dividing daughter cells, requires coordination of myosin II function, membrane trafficking, and central spindle organization. Most known regulators act during late cytokinesis; a few, including the myosin II-binding proteins anillin and supervillin, act earlier. Anillin's role in scaffolding the membrane cortex with the central spindle is well established, but the mechanism of supervillin action is relatively uncharacterized. We show here that two regions within supervillin affect cell division: residues 831-1281, which bind central spindle proteins, and residues 1-170, which bind the myosin II heavy chain (MHC) and the long form of myosin light-chain kinase. MHC binding is required to rescue supervillin deficiency, and mutagenesis of this site creates a dominant-negative phenotype. Supervillin concentrates activated and total myosin II at the furrow, and simultaneous knockdown of supervillin and anillin additively increases cell division failure. Knockdown of either protein causes mislocalization of the other, and endogenous anillin increases upon supervillin knockdown. Proteomic identification of interaction partners recovered using a high-affinity green fluorescent protein nanobody suggests that supervillin and anillin regulate the myosin II and actin cortical cytoskeletons through separate pathways. We conclude that supervillin and anillin play complementary roles during vertebrate cytokinesis.
authors.affiliation: Program in Cell and Developmental Dynamics, Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, MA 01655 Laboratory of Cellular and Structural Biology, Rockefeller University, New York, NY 10065 Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, Rockefeller University, New York, NY 10065 Proteomics and Mass Spectrometry Facility, University of Massachusetts Medical School, Shrewsbury, MA 01545.
authors.forename: Tara C
authors.initials: TC
authors.lastname: Smith
authors.forename: Peter C
authors.initials: PC
authors.lastname: Fridy
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Li
authors.forename: Shruti
authors.initials: S
authors.lastname: Basil
authors.forename: Sneha
authors.initials: S
authors.lastname: Arjun
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Friesen
authors.forename: John
authors.initials: J
authors.lastname: Leszyk
authors.forename: Brian T
authors.initials: BT
authors.lastname: Chait
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Luna
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003285:Contractile Proteins / Q000378:metabolism*; D048749:Cytokinesis; D003599:Cytoskeleton / Q000378:metabolism; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism*; D016296:Mutagenesis; D024462:Myosin Type II / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D017354:Point Mutation; D011485:Protein Binding; D021381:Protein Transport; D017384:Sequence Deletion; D015398:Signal Transduction
chemical_list: D003285:Contractile Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; C109413:SVIL protein, human; C095886:anillin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D009219:Myosin-Light-Chain Kinase; D024462:Myosin Type II